Fredrik Lanner1, Marcus Sohl, Filip Farnebo. 1. Department of Cell and Molecular Biology, Karolinska Institutet, Box 285, SE-17177 Stockholm, Sweden. fredrik.lanner@ki.se
Abstract
OBJECTIVE: The aim of this work was to develop a mouse embryonic stem (ES) cell system addressing the early specification of the developing vasculature into functional arteries and veins. METHODS AND RESULTS: ES cells were differentiated 4 days on collagen-type IV coated dishes to obtain Flk1+ endothelial precursors. Sub-culture of these precursors for additional 4 days robustly generated, in a VEGF dose-dependent manner, mature endothelial cells. Arterial marker genes were specifically expressed in cultures differentiated with high VEGF concentration whereas the venous marker gene COUP-TFII was upregulated in endothelial cells induced through low and intermediate VEGF concentrations. This VEGF-dependent arterialization could be blocked by inhibition of Notch resulting in an arterial to venous fate switch. Functional and morphological studies, ie, measurement of sprout length, pericyte recruitment, and interleukin-I-induced leukocyte adhesion, further confirmed their arterial and venous identity. CONCLUSIONS: We conclude that endothelial cells with distinct molecular, morphological, and functional characteristics of arteries and veins can be derived through in vitro differentiation of ES cells in a VEGF dose-dependent and Notch-regulated manner.
OBJECTIVE: The aim of this work was to develop a mouse embryonic stem (ES) cell system addressing the early specification of the developing vasculature into functional arteries and veins. METHODS AND RESULTS: ES cells were differentiated 4 days on collagen-type IV coated dishes to obtain Flk1+ endothelial precursors. Sub-culture of these precursors for additional 4 days robustly generated, in a VEGF dose-dependent manner, mature endothelial cells. Arterial marker genes were specifically expressed in cultures differentiated with high VEGF concentration whereas the venous marker gene COUP-TFII was upregulated in endothelial cells induced through low and intermediate VEGF concentrations. This VEGF-dependent arterialization could be blocked by inhibition of Notch resulting in an arterial to venous fate switch. Functional and morphological studies, ie, measurement of sprout length, pericyte recruitment, and interleukin-I-induced leukocyte adhesion, further confirmed their arterial and venous identity. CONCLUSIONS: We conclude that endothelial cells with distinct molecular, morphological, and functional characteristics of arteries and veins can be derived through in vitro differentiation of ES cells in a VEGF dose-dependent and Notch-regulated manner.
Authors: Jinjoo Kang; Jaehyuk Yoo; Sunju Lee; Wanli Tang; Berenice Aguilar; Swapnika Ramu; Inho Choi; Hasan H Otu; Jay W Shin; G Paolo Dotto; Chester J Koh; Michael Detmar; Young-Kwon Hong Journal: Blood Date: 2010-03-29 Impact factor: 22.113
Authors: Melinda Hajdu; Aernout Luttun; Beatriz Pelacho; Terry C Burns; Lucas Chase; María Gutiérrez-Pérez; Yuehua Jiang; Todd Lenvik; Virág Vas; Ferenc Uher; Anna Sebestyén; Catherine Verfaillie Journal: Pathol Oncol Res Date: 2007-12-25 Impact factor: 3.201
Authors: Mark P White; Abdul J Rufaihah; Lei Liu; Yohannes T Ghebremariam; Kathryn N Ivey; John P Cooke; Deepak Srivastava Journal: Stem Cells Date: 2013-01 Impact factor: 6.277